Cumulative evidence suggests that O-linkedβ-N-acetylglucosaminylation(OGlcNAcylation)plays an important regulatory role in pathophysiological processes.Although the regulatory mechanisms of O-GlcNAcylation in tumors ...Cumulative evidence suggests that O-linkedβ-N-acetylglucosaminylation(OGlcNAcylation)plays an important regulatory role in pathophysiological processes.Although the regulatory mechanisms of O-GlcNAcylation in tumors have been gradually elucidated,the potential mechanisms of O-GlcNAcylation in bone metabolism,particularly,in the osteogenic differentiation of bone marrow mesenchymal stromal cells(BMSCs)remains unexplored.In this study,the literature related to O-GlcNAcylation and BMSC osteogenic differentiation was reviewed,assuming that it could trigger more scholars to focus on research related to OGlcNAcylation and bone metabolism and provide insights into the development of novel therapeutic targets for bone metabolism disorders such as osteoporosis.展开更多
Mesenchymal stem cells(MSCs)can differentiate into various tissue cell types including bone,adipose,cartilage,and muscle.Among those,osteogenic differentiation of MSCs has been widely explored in many bone tissue engi...Mesenchymal stem cells(MSCs)can differentiate into various tissue cell types including bone,adipose,cartilage,and muscle.Among those,osteogenic differentiation of MSCs has been widely explored in many bone tissue engineering studies.Moreover,the conditions and methods of inducing osteogenic differentiation of MSCs are continuously advancing.Recently,with the gra-dual recognition of adipokines,the research on their involvement in different pathophysiological processes of the body is also deepening including lipid metabolism,inflammation,immune regulation,energy disorders,and bone homeostasis.At the same time,the role of adipokines in the osteogenic differentiation of MSCs has been gradually described more completely.Therefore,this paper reviewed the evidence of the role of adipokines in the osteogenic differentiation of MSCs,emphasizing bone formation and bone regeneration.展开更多
Receptor tyrosine kinase-like orphan receptor 2(ROR2)has a vital role in osteogenesis.However,the mechanism underlying the regulation of ROR2 in osteogenic differentiation is still poorly comprehended.A previous study...Receptor tyrosine kinase-like orphan receptor 2(ROR2)has a vital role in osteogenesis.However,the mechanism underlying the regulation of ROR2 in osteogenic differentiation is still poorly comprehended.A previous study by our research group showed that a novel compound heterozygous ROR2 variation accounted for the autosomal recessive Robinow syndrome(ARRS).This study attempted to explore the impact of the ROR2:c.904C>T variant specifically on the osteogenic differentiation of BMSCs.Methods:Coimmunoprecipitation(CoIP)-western blotting was carried out to identify the interaction between ROR2 and Wnt5a.Double-immunofluorescence staining was used for determining the expressions and co-localization of ROR2 and Wnt5a in bone marrow mesenchymal stem cells(BMSCs).Western blot(WB)analysis and quantitative reverse transcription polymerase chain reaction(RT-qPCR)were conducted to identify the expression levels of ROR2 in the BMSCs transfected with LV-shROR2 or LV-ROR2-c.904C>T.The alkaline phosphatase(ALP)activity was detected,and Alizarin Red S staining was done for evaluating the osteogenic differentiation of BMSCs.RT-qPCR was employed to identify the expression of the sphingomyelin synthase 1(SMS1)mRNA in the BMSCs transfected with LV-shROR2 or LV-ROR2-c.904C>T and the mRNA expression levels of Runt-related transcription factor 2(RUNX2),osteocalcin(OCN),and osteopontin(OPN).WB was performed to confirm the protein expressions of extracellular regulated protein kinases1(ERK),P-ERK,Smad family member1/5/8(Smad1/5/8),P-Smad1/5/8,P-P38,P38,RUNX2,OCN,and OPN in the BMSCs transfected with LV-shROR2/LV-ROR2-c.904C>T and sphingomyelin(SM).Results:The ROR2:c.904C>T mutant altered the subcellular localization of the ROR2 protein,which caused an impaired interaction between ROR2 and Wnt5a.The depletion of ROR2 restricted the osteogenic differentiation capability of BMSCs and downregulated the expression of SMS1.SM treatment could reverse the inhibition of osteoblastic differentiation in ROR2-depleted BMSCs.Conclusion:The findings of this work revealed that the ROR2:c.904C>T variant led to the loss of function of ROR2,which impaired the interaction between ROR2 and Wnt5a and also controlled the osteogenic differentiation capability of BMSCs.Furthermore,SM was revealed to be engaged in the osteoblastic differentiation of BMSCs regulated by ROR2,which renders SM a potential target in the therapy for ARRS.展开更多
BACKGROUND The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells(PSCs)into osteoblasts or chondrocytes;however,the underlying mechanisms remain unclear.AIM...BACKGROUND The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells(PSCs)into osteoblasts or chondrocytes;however,the underlying mechanisms remain unclear.AIM To determine the effect of hypoxia on PSCs,and the expression of microRNA-584-5p(miR-584-5p)and RUNX family transcription factor 2(RUNX2)in PSCs was modulated to explore the impact of the miR-584-5p/RUNX2 axis on hypoxiainduced osteogenic differentiation of PSCs.METHODS In this study,we isolated primary mouse PSCs and stimulated them with hypoxia,and the characteristics and functional genes related to PSC osteogenic differentiation were assessed.Constructs expressing miR-584-5p and RUNX2 were established to determine PSC osteogenic differentiation.RESULTS Hypoxic stimulation induced PSC osteogenic differentiation and significantly increased calcified nodules,intracellular calcium ion levels,and alkaline phosphatase(ALP)activity in PSCs.Osteogenic differentiation-related factors such as RUNX2,bone morphogenetic protein 2,hypoxia-inducible factor 1-alpha,and ALP were upregulated;in contrast,miR-584-5p was downregulated in these cells.Furthermore,upregulation of miR-584-5p significantly inhibited RUNX2 expression and hypoxia-induced PSC osteogenic differentiation.RUNX2 was the target gene of miR-584-5p,antagonizing miR-584-5p inhibition in hypoxia-induced PSC osteogenic differentiation.CONCLUSION Our study showed that the interaction of miR-584-5p and RUNX2 could mediate PSC osteogenic differentiation induced by hypoxia.展开更多
ALKBH1 was recently discovered as a demethylase for DNA N^6-methyladenine(N6-m A), a new epigenetic modification, and interacts with the core transcriptional pluripotency network of embryonic stem cells.However, the r...ALKBH1 was recently discovered as a demethylase for DNA N^6-methyladenine(N6-m A), a new epigenetic modification, and interacts with the core transcriptional pluripotency network of embryonic stem cells.However, the role of ALKBH1 and DNA N6-m A in regulating osteogenic differentiation is largely unknown.In this study, we demonstrated that the expression of ALKBH1 in human mesenchymal stem cells(MSCs)was upregulated during osteogenic induction. Knockdown of ALKBH1 increased the genomic DNA N6-m A levels and significantly reduced the expression of osteogenic-related genes, alkaline phosphatase activity, and mineralization. ALKBH1-depleted MSCs also exhibited a restricted capacity for bone formation in vivo.By contrast, the ectopic overexpression of ALKBH1 enhanced osteoblastic differentiation. Mechanically,we found that the depletion of ALKBH1 resulted in the accumulation of N6-m A on the promoter region of ATF4, which subsequently silenced ATF4 transcription. In addition, restoring the expression of ATP by adenovirus-mediated transduction successfully rescued osteogenic differentiation. Taken together, our results demonstrate that ALKBH1 is indispensable for the osteogenic differentiation of MSCs and indicate that DNA N6-m A modifications area new mechanism for the epigenetic regulation of stem cell differentiation.展开更多
Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in me...Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.展开更多
A facile modification strategy is developed to promote the proliferation and osteogenic differentiation of rat bone marrow stromal cells(rBMSCs)through deposition of a bioactive calcium silicate(CS)coating on the poro...A facile modification strategy is developed to promote the proliferation and osteogenic differentiation of rat bone marrow stromal cells(rBMSCs)through deposition of a bioactive calcium silicate(CS)coating on the porous surface of poly(ether-ether-ketone)(PEEK)with the assistance of poly(dopamine)(PDA).The porous structures are etched on the surface of PEEK after sulfonation treatment.A poly(dopamine)layer is coated on the porous surface of the sulfonated PEEK(SPEEK),which provides anchoring groups for the subsequent deposition of the CS layer.Results show that the CS coating on the porous surface of SPEEK significantly improve the hydrophilicity and biomineralization formation of hydroxyapatite.Compared with PEEK,SPEEK-PDA-CS displays higher bioactivity to promote the proliferation and osteogenic differentiation of rBMSCs,including the increase of ALP activity and formation of calcium nodules,the expression of osteogenic differentiation-related genes.These results are beneficial to extending clinical applications of PEEK-based implants for bone tissue repair and orthopedic surgery.展开更多
The present study was designed to investigate the role of estrogen receptorα(ERα)in biaxial tensile strain(BTS)regulated osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells(rBMSCs).rBMSCs we...The present study was designed to investigate the role of estrogen receptorα(ERα)in biaxial tensile strain(BTS)regulated osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells(rBMSCs).rBMSCs were derived fromrats and overexpressed ERα.The rBMSCs were subjected to BTS at 1Hz with a strain of 2%for 4 h per day,3 days,with or without ERαinhibitor ICI 182,780(ICI).Then,bone mineralization was performed by Alizarin Red Staining.The markers of osteogenic differentiation and downstream Wnt3a/β-catenin signaling were detected by western blotting.Results showed that BTS enhanced the osteogenic differentiation of rBMSCs,increased protein expression levels of alkaline phosphatase(ALP),runt-related transcription factor 2(Runx2),collagen type I(Col I)and osteocalcin(OCN),and it increased the protein expression levels of estrogen receptor(ER)α(ERα),Wnt3a,andβ-catenin.BTS The activated Wnt3a/β-catenin signaling pathway induced by BTS was abolished by ICI 182,780(ICI).In addition,overexpressing ERαin rBMSCs promoted the osteogenic differentiation by BTS.Taken together,BTS induced osteogenic differentiation of rBMSCs via the ERαand downstream canonical Wnt3a/β-catenin pathway.展开更多
Human mesenchymal stem cells(hMSC)can be expanded in vitro and differentiated towards osteogenic,chondrogenic or adipogenic lineages,making them an attractive source for tissue engineering and regenerative medicine.Ch...Human mesenchymal stem cells(hMSC)can be expanded in vitro and differentiated towards osteogenic,chondrogenic or adipogenic lineages,making them an attractive source for tissue engineering and regenerative medicine.Chitinase-like-proteins(CLPs)belong to the family 18 glycosyl hydrolases and are believed to play a role in inflammation and tissue remodelling.The aim of this study was to determine the effect of the aminosugar glucosamine on the expression of the CLP YKL-40 during osteogenic differentiation of hMSC.Glucosamine did not affect multipotency of hMSC nor proliferation rate of undifferentiated hMSC.YKL-40 was expressed during both expansion of undifferentiated hMSC and during osteogenic differentiation.A slight but non-significant increase in YKL-40 expression was observed with glucosamine,accompanied by a pH-dependent delay in mineralization.However,glucosamine induced higher expression of osteogenic marker genes.展开更多
Nanotopographical features are found to have significant effects on bone behavior. In the present study, nanoporous aluminas with different pore sizes (20, 100 and 200 nm in diameter), were evaluated for their osteoin...Nanotopographical features are found to have significant effects on bone behavior. In the present study, nanoporous aluminas with different pore sizes (20, 100 and 200 nm in diameter), were evaluated for their osteoinductive and drug eluting properties. W20-17 marrow stromal cells were seeded on nanoporous alumina with and without the addition of BMP-2. Although cell proliferation was not affected by pore size, osteogenic differentiation was 200 nm as compared to 20 and 100 nm pores induced higher alkaline phosphatase activity (ALP) and osteocalcin expression levels, thus indicating osteoblastic differentiation. Cell morphology revealed that cells cultured on 20 nm pores adopted a rounded shape, while larger pores (200 nm) elicited an elongated morphology. Furthermore, ALP expression levels were consistently higher on BMP-2 loaded nanoporous alumina surfaces compared to unloaded surfaces, indicating that not only is nanoporous alumina osteoinductive, but also has the potential to be used as a drug eluting bone-implant coating.展开更多
Objective To study the effects of combination of bone morphogenetic protein-2 (BMP-2) and strontium chloride on proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells(hUCM-SCs)in v...Objective To study the effects of combination of bone morphogenetic protein-2 (BMP-2) and strontium chloride on proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells(hUCM-SCs)in vitro culture.展开更多
Periodontal diseases are infectious diseases that are characterized by progressive damage to dental support tissue.The major goal of periodontal therapy is to regenerate the periodontium destroyed by periodontal disea...Periodontal diseases are infectious diseases that are characterized by progressive damage to dental support tissue.The major goal of periodontal therapy is to regenerate the periodontium destroyed by periodontal diseases.Human periodontal ligament(PDL)tissue possesses periodontal regenerative properties,and periodontal ligament stem cells(PDLSCs)with the capacity for osteogenic differentiation show strong potential in clinical application for periodontium repair and regeneration.Noncoding RNAs(ncRNAs),which include a substantial portion of poly-A tail mature RNAs,are considered“transcriptional noise.”Recent studies show that ncRNAs play a major role in PDLSC differentiation;therefore,exploring how ncRNAs participate in the osteogenic differentiation of PDLSCs may help to elucidate the underlying mechanism of the osteogenic differentiation of PDLSCs and further shed light on the potential of stem cell transplantation for periodontium regeneration.In this review paper,we discuss the history of PDLSC research and highlight the regulatory mechanism of ncRNAs in the osteogenic differentiation of PDLSCs.展开更多
Objective To observe the expression profiles of osteoblast-related genes in human mesenchymal stem cells(MSCs) derived from bone marrow during osteogenic differentiation. Methods MSCs were induced to differentiate wit...Objective To observe the expression profiles of osteoblast-related genes in human mesenchymal stem cells(MSCs) derived from bone marrow during osteogenic differentiation. Methods MSCs were induced to differentiate with MSC osteogenic differentiation medium for 7, 14, 21 and 28 days respectively. Alizarin Red staining was used to detect matrix mineralization. Expression of osteoblast-related genes, including osteocalcin, osteopontin, Runt-related transcription factor 2(Runx2), alkaline phosphatase and collagen type Ⅰ, was assessed with quantitative reverse transcription-polymerase chain reaction. Results On day 14 after induction of differentiation, cells were stained positively with Alizarin Red. The expression levels of these genes exhibited an upward trend as induction time was prolonged. Exposure to osteogenic differentiation medium less than 21 days did not significantly induce osteocalcin expression; osteocalcin expression levels in the differentiated cells induced for 21 and 28 days were 1.63 and 2.46 times as high as the undifferentiated cells respectively(all P<0.05). Stimulation with MSC osteogenic differentiation medium over 14 days significantly enhanced bone marrow-derived MSCs to express osteopontin and Runx2 genes(all P<0.05). Osteogenic differentiation medium could significantly induce the expressions of alkaline phosphatase and collagen type Ⅰgenes(all P<0.05). Their expressions reached the peak levels on day 21, which were increased more than 4- and 3-fold respectively. Conclusion Human bone marrow-derived MSCs could exhibit the sequential expression pattern of osteoblast marker genes during osteogenic differentiation in vitro.展开更多
Sites of implantation with compromised biology may be unable to achieve the required level of angiogenic and osteogenic regeneration. The specific function and contribution of different cell types to the formation of ...Sites of implantation with compromised biology may be unable to achieve the required level of angiogenic and osteogenic regeneration. The specific function and contribution of different cell types to the formation of prevascularized, osteogenic networks in co-culture remains unclear. To determine how bone marrow-derived mesenchymal stromal cells(BMSCs) and endothelial cells(ECs) contribute to cellular proangiogenic differentiation, we analysed the differentiation of BMSCs and ECs in standardized monolayer, Transwell and co-cultures. BMSCs were derived from the iliac bone marrow of five patients,characterized and differentiated in standardized monolayers, permeable Transwells and co-cultures with human umbilical vein ECs(HUVECs). The expression levels of CD31, von Willebrand factor, osteonectin(ON) and Runx2 were assessed by quantitative reverse transcriptase polymerase chain reaction. The protein expression of alkaline phosphatase, ON and CD31 was demonstrated via histochemical and immunofluorescence analysis. The results showed that BMSCs and HUVECs were able to retain their lineage-specific osteogenic and angiogenic differentiation in direct and indirect co-cultures. In addition, BMSCs demonstrated a supportive expression of angiogenic function in co-culture, while HUVEC was able to improve the expression of osteogenic marker molecules in BMSCs.展开更多
Retaining or improving periodontal ligament (PDL) function is crucial for restoring periodontal defects. The aim of this study was to evaluate the physiological effects of low-power laser irradiation (LPLI) on the pro...Retaining or improving periodontal ligament (PDL) function is crucial for restoring periodontal defects. The aim of this study was to evaluate the physiological effects of low-power laser irradiation (LPLI) on the proliferation and osteogenic differentiation of human PDL (hPDL) cells. Cultured hPDL cells were irradiated (660 nm) daily with doses of 0, 1, 2 or 4 J·cm-2 . Cell proliferation was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, and the effect of LPLI on osteogenic differentiation was assessed by Alizarin Red S staining and alkaline phosphatase (ALP) activity. Additionally, osteogenic marker gene expression was confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR). Our data showed that LPLI at a dose of 2 J·cm-2 significantly promoted hPDL cell proliferation at days 3 and 5. In addition, LPLI at energy doses of 2 and 4 J·cm-2 showed potential osteogenic capacity, as it stimulated ALP activity, calcium deposition, and osteogenic gene expression. We also showed that cyclic adenosine monophosphate (cAMP) is a critical regulator of the LPLI-mediated effects on hPDL cells. This study shows that LPLI can promote the proliferation and osteogenic differentiation of hPDL cells. These results suggest the potential use of LPLI in clinical applications for periodontal tissue regeneration.展开更多
Chronic high glucose(HG) plays a crucial role in the pathogenesis of diabetes-induced osteoporosis by inhibiting the differentiation and proliferation of osteoblasts. This study aims to examine the role of E26 transfo...Chronic high glucose(HG) plays a crucial role in the pathogenesis of diabetes-induced osteoporosis by inhibiting the differentiation and proliferation of osteoblasts. This study aims to examine the role of E26 transformation-specific 1(ETS1) in the inhibition of osteoblast differentiation and proliferation caused by chronic HG, as well as the underlying mechanism. Chronic HG treatment downregulated ETS1 expression and inhibited differentiation and proliferation of MC3 T3-E1 cells. Downregulation of ETS1 expression inhibited the differentiation and proliferation of MC3 T3-E1 cells under normal glucose conditions, and ETS1 overexpression attenuated the damage to cells exposed to chronic HG. In addition, ETS1 overexpression reversed the decrease in runt-related transcription factor 2(Runx2) expression in MC3 T3-E1 cells treated with chronic HG. Using chromatin immunoprecipitation(ChIP) and luciferase reporter assays, we confirmed that ETS1 directly bound to and increased the activity of the Runx2 promoter. In summary, our study suggested that ETS1 was involved in the inhibitory effect of chronic HG on osteogenic differentiation and proliferation and may be a potential therapeutic target for diabetes-induced osteoporosis.展开更多
Poly (lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol)(PLGA-[ASP-PEG]) scaffold materials were linked with a novel nonviral vector (K)16GRGDSPC through cross linker Sulfo- LC-SPDP to construct a new ty...Poly (lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol)(PLGA-[ASP-PEG]) scaffold materials were linked with a novel nonviral vector (K)16GRGDSPC through cross linker Sulfo- LC-SPDP to construct a new type of nonviral gene transfer system. Eukaryotic expressing vector containing transforming growth factor beta 1 (pcDNA3-TGFβ1) was encapsulated by the system. Bone marrow stromal cells (BMSCs) obtained from rabbit were cultured on PLGA-[ASP-PEG] modified by (K)16GRGDSPC and TGF-β1 gene and PLGA-[ASP-PEG] modified by (K)16GRGDSPC and empty vector pcDNA3 as control. The expressions of osteogenic makers of the BMSCs cultured on the TGF-β1 gene-activated scaffold materials were found significantly higher than those of the control group (P<0.05). A brand-new way was provided for regulating seed cells to directionally differentiate into osteoblasts for bone defect restoration in bone tissue engineering.展开更多
Bone tissue engineering, aiming at developing bone substitutes for repair and regeneration of bone defects instead of using autologous bone grafts,has attracted wide attention in the field of tissue engineering and re...Bone tissue engineering, aiming at developing bone substitutes for repair and regeneration of bone defects instead of using autologous bone grafts,has attracted wide attention in the field of tissue engineering and regenerative medicine.Developing biomimetic biomaterial scaffolds able to regulate osteogenic differentiation of stem cells could be a promising strategy to improve the therapeutic efficacy.In this study, electrospun composite nanofibers of hydroxyapatite / collagen / chitosan( HAp / Col / CTS)resembling the fibrous nanostructure and constituents of the hierarchically organized natural bone,were prepared to investigate their capacity for promoting bone mesenchymal stem cells( BMSCs)to differentiate into the osteogenic lineage in the absence and presence of the osteogenic supplementation, respectively.Cell morphology,proliferation and quantified specific osteogenic protein expression on the electrospun HAp / Col / CTS scaffolds were evaluated in comparison with different controls including electrospun nanofibrous CTS,HAp / CTS and tissue culture plate.Our results showed that the nanofibrous HAp / Col / CTS scaffolds supported better spreading and proliferation of the BMSCs than other substrates( P < 0.01).Expressions of osteogenesis protein markers,alkaline phosphatase( ALP) and Col,were significantly upregulated on the HAp / Col / CTS than those on the CTS( P < 0.01) and HAp /CTS( P < 0.05) scaffolds in the absence of the osteogenic supplementation.Moreover,presence of osteogenic supplementation also proved to enhance osteogenic differentiation of BMSCs on HAp /Col / CTS scaffolds, indicative of a synergistic effect.This study highlights the potential of BMSCs / HAp / Col / CTS cell-scaffold system for functional bone repair and regeneration applications.展开更多
The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human...The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) phenotypic characterization, 2) The in-vitro osteogenic differentiation potential of hUCMSCs, 3) The cytotoxicity of gelatin solvent to hUCMSCs using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. As a result, through characterization of hUCMSCs, the majority of target cells expressed specific MSCs markers, Cellular Differentiation (CD)73, smaller number of subpopulation expressed CD90 with only minimal subpopulation expressed CD105 and all negative MSCs markers. Osteoblastic differentiation was found in a significantly high number of cells when in vitro osteogenic differentiation of hUCMSCs with Alizarin Red staining was done. Biocompatibility analysis using the MTT assay showed that gelatin solvent and Alpha modification of minimum essential medium Eagle (α-MEM) was non-toxic for hUCMSCs in vitro. The study concluded that hUCMSCs isolated from human umbilical cord was capable of undergoing in vitro osteogenesis, indicating its potential as allogeneic stem cells for clinical application in bone tissue engineering.展开更多
Objective:To investigate the effects and mechanisms of Polygonatum sibiricum polysaccharide(PSP)on osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods:The mouse BMSCs were cultured and indu...Objective:To investigate the effects and mechanisms of Polygonatum sibiricum polysaccharide(PSP)on osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods:The mouse BMSCs were cultured and induced in osteoblast medium(OBM)containing finalconcentrations(0 mg/L,12.5 mg/L,25 mg/L,and 50 mg/L)of PSP.The proliferation and cytotoxicity of BMSCs were detected by MTT assay.Alkaline phosphatase(ALP)and Alizarin red S staining were performed after 7 days' ossification-inducing culture.The mRNA expressions of ALP,Runx2 and osteocalcin(OCN)were determined by fluorescence quantitative PCR(qPCR).The mRNA and protein expressions of Tafazzin(TAZ)(a key effector of Hippo pathway)were measured by qPCR and western blotting,respectively.Results:PSP was non-cytotoxicwithin the dose range of 12.5-50 mg/L and had no impact on the proliferation of BMSCs.The activity of ALP,the intensity of ALP staining,and the formation of mineralized nodules were increased by PSP treatment(25 and50 mg/L)(P<0.01).Moreover,administration of 25 mg/L PSP significantly enhanced the mRNA levels of osteoblastic differentiation makers ALP,Runx2 and OCN as well as the mRNA and protein expressions of TAZ(P<0.01).Conclusion:PSP could promote osteogenic differentiation of BMSCs,and the mechanisms might be related to the activation of TAZ in the Hippo signaling pathway.展开更多
文摘Cumulative evidence suggests that O-linkedβ-N-acetylglucosaminylation(OGlcNAcylation)plays an important regulatory role in pathophysiological processes.Although the regulatory mechanisms of O-GlcNAcylation in tumors have been gradually elucidated,the potential mechanisms of O-GlcNAcylation in bone metabolism,particularly,in the osteogenic differentiation of bone marrow mesenchymal stromal cells(BMSCs)remains unexplored.In this study,the literature related to O-GlcNAcylation and BMSC osteogenic differentiation was reviewed,assuming that it could trigger more scholars to focus on research related to OGlcNAcylation and bone metabolism and provide insights into the development of novel therapeutic targets for bone metabolism disorders such as osteoporosis.
基金the Changzhou Science&Technology Program,No.CJ20210104,CJ20220120,and CJ20210005Qinghai Province Health System Guidance Plan Project,No.2022-wjzdx-106+1 种基金Young Talent Development Plan of Changzhou Health commission,No.CZQM2020059Top Talent of Changzhou“The 14th Five-Year Plan”High-Level Health Talents Training Project,No.2022CZBJ059 and 2022CZBJ061.
文摘Mesenchymal stem cells(MSCs)can differentiate into various tissue cell types including bone,adipose,cartilage,and muscle.Among those,osteogenic differentiation of MSCs has been widely explored in many bone tissue engineering studies.Moreover,the conditions and methods of inducing osteogenic differentiation of MSCs are continuously advancing.Recently,with the gra-dual recognition of adipokines,the research on their involvement in different pathophysiological processes of the body is also deepening including lipid metabolism,inflammation,immune regulation,energy disorders,and bone homeostasis.At the same time,the role of adipokines in the osteogenic differentiation of MSCs has been gradually described more completely.Therefore,this paper reviewed the evidence of the role of adipokines in the osteogenic differentiation of MSCs,emphasizing bone formation and bone regeneration.
基金funded by the Project Funded by China Postdoctoral Science Foundation(No.2022T150445)the Beijing Hospitals Authority Youth Programme(No.QML20211401)+1 种基金the Young Talent Foundation of PLA General Hospital(2019-YQPY-002)Beijing Nova Program(Z201100006820057).
文摘Receptor tyrosine kinase-like orphan receptor 2(ROR2)has a vital role in osteogenesis.However,the mechanism underlying the regulation of ROR2 in osteogenic differentiation is still poorly comprehended.A previous study by our research group showed that a novel compound heterozygous ROR2 variation accounted for the autosomal recessive Robinow syndrome(ARRS).This study attempted to explore the impact of the ROR2:c.904C>T variant specifically on the osteogenic differentiation of BMSCs.Methods:Coimmunoprecipitation(CoIP)-western blotting was carried out to identify the interaction between ROR2 and Wnt5a.Double-immunofluorescence staining was used for determining the expressions and co-localization of ROR2 and Wnt5a in bone marrow mesenchymal stem cells(BMSCs).Western blot(WB)analysis and quantitative reverse transcription polymerase chain reaction(RT-qPCR)were conducted to identify the expression levels of ROR2 in the BMSCs transfected with LV-shROR2 or LV-ROR2-c.904C>T.The alkaline phosphatase(ALP)activity was detected,and Alizarin Red S staining was done for evaluating the osteogenic differentiation of BMSCs.RT-qPCR was employed to identify the expression of the sphingomyelin synthase 1(SMS1)mRNA in the BMSCs transfected with LV-shROR2 or LV-ROR2-c.904C>T and the mRNA expression levels of Runt-related transcription factor 2(RUNX2),osteocalcin(OCN),and osteopontin(OPN).WB was performed to confirm the protein expressions of extracellular regulated protein kinases1(ERK),P-ERK,Smad family member1/5/8(Smad1/5/8),P-Smad1/5/8,P-P38,P38,RUNX2,OCN,and OPN in the BMSCs transfected with LV-shROR2/LV-ROR2-c.904C>T and sphingomyelin(SM).Results:The ROR2:c.904C>T mutant altered the subcellular localization of the ROR2 protein,which caused an impaired interaction between ROR2 and Wnt5a.The depletion of ROR2 restricted the osteogenic differentiation capability of BMSCs and downregulated the expression of SMS1.SM treatment could reverse the inhibition of osteoblastic differentiation in ROR2-depleted BMSCs.Conclusion:The findings of this work revealed that the ROR2:c.904C>T variant led to the loss of function of ROR2,which impaired the interaction between ROR2 and Wnt5a and also controlled the osteogenic differentiation capability of BMSCs.Furthermore,SM was revealed to be engaged in the osteoblastic differentiation of BMSCs regulated by ROR2,which renders SM a potential target in the therapy for ARRS.
基金Supported by Sailing Program of Naval Medical University,Program of Shanghai Hongkou District Health Commission,No.2202-27Special Funds for Activating Scientific Research of Shanghai Fourth People’s Hospital,No.sykyqd05801.
文摘BACKGROUND The hypoxic environment during bone healing is important in regulating the differentiation of periosteal stem cells(PSCs)into osteoblasts or chondrocytes;however,the underlying mechanisms remain unclear.AIM To determine the effect of hypoxia on PSCs,and the expression of microRNA-584-5p(miR-584-5p)and RUNX family transcription factor 2(RUNX2)in PSCs was modulated to explore the impact of the miR-584-5p/RUNX2 axis on hypoxiainduced osteogenic differentiation of PSCs.METHODS In this study,we isolated primary mouse PSCs and stimulated them with hypoxia,and the characteristics and functional genes related to PSC osteogenic differentiation were assessed.Constructs expressing miR-584-5p and RUNX2 were established to determine PSC osteogenic differentiation.RESULTS Hypoxic stimulation induced PSC osteogenic differentiation and significantly increased calcified nodules,intracellular calcium ion levels,and alkaline phosphatase(ALP)activity in PSCs.Osteogenic differentiation-related factors such as RUNX2,bone morphogenetic protein 2,hypoxia-inducible factor 1-alpha,and ALP were upregulated;in contrast,miR-584-5p was downregulated in these cells.Furthermore,upregulation of miR-584-5p significantly inhibited RUNX2 expression and hypoxia-induced PSC osteogenic differentiation.RUNX2 was the target gene of miR-584-5p,antagonizing miR-584-5p inhibition in hypoxia-induced PSC osteogenic differentiation.CONCLUSION Our study showed that the interaction of miR-584-5p and RUNX2 could mediate PSC osteogenic differentiation induced by hypoxia.
基金supported by grants from the National Natural Science Foundation of China (No.81271178 and 81470777)
文摘ALKBH1 was recently discovered as a demethylase for DNA N^6-methyladenine(N6-m A), a new epigenetic modification, and interacts with the core transcriptional pluripotency network of embryonic stem cells.However, the role of ALKBH1 and DNA N6-m A in regulating osteogenic differentiation is largely unknown.In this study, we demonstrated that the expression of ALKBH1 in human mesenchymal stem cells(MSCs)was upregulated during osteogenic induction. Knockdown of ALKBH1 increased the genomic DNA N6-m A levels and significantly reduced the expression of osteogenic-related genes, alkaline phosphatase activity, and mineralization. ALKBH1-depleted MSCs also exhibited a restricted capacity for bone formation in vivo.By contrast, the ectopic overexpression of ALKBH1 enhanced osteoblastic differentiation. Mechanically,we found that the depletion of ALKBH1 resulted in the accumulation of N6-m A on the promoter region of ATF4, which subsequently silenced ATF4 transcription. In addition, restoring the expression of ATP by adenovirus-mediated transduction successfully rescued osteogenic differentiation. Taken together, our results demonstrate that ALKBH1 is indispensable for the osteogenic differentiation of MSCs and indicate that DNA N6-m A modifications area new mechanism for the epigenetic regulation of stem cell differentiation.
基金supported by grants from the National Natural Science Foundation of China(82071150,82170934,81870743,8190104 and 82171001)。
文摘Human adipose-derived stem cells(hASCs)are a promising cell type for bone tissue regeneration.Circular RNAs(circRNAs)have been shown to play a critical role in regulating various cell differentiation and involve in mesenchymal stem cell osteogenesis.However,how circRNAs regulate hASCs in osteogenesis is still unclear.Herein,we found circ_0003204 was significantly downregulated during osteogenic differentiation of hASCs.Knockdown of circ_0003204 by si RNA or overexpression by lentivirus confirmed circ_0003204 could negatively regulate the osteogenic differentiation of hASCs.We performed dual-luciferase reporting assay and rescue experiments to verify circ_0003204 regulated osteogenic differentiation via sponging miR-370-3p.We predicted and confirmed that miR-370-3p had targets in the 3′-UTR of HDAC4 m RNA.The following rescue experiments indicated that circ_0003204 regulated the osteogenic differentiation of hASCs via miR-370-3p/HDAC4 axis.Subsequent in vivo experiments showed the silencing of circ_0003204 increased the bone formation and promoted the expression of osteogenic-related proteins in a mouse bone defect model,while overexpression of circ_0003204 inhibited bone defect repair.Our findings indicated that circ_0003204 might be a promising target to promote the efficacy of hASCs in repairing bone defects.
基金Supported by the National Key Research&Development Program of China(No.2018YFB1105702)。
文摘A facile modification strategy is developed to promote the proliferation and osteogenic differentiation of rat bone marrow stromal cells(rBMSCs)through deposition of a bioactive calcium silicate(CS)coating on the porous surface of poly(ether-ether-ketone)(PEEK)with the assistance of poly(dopamine)(PDA).The porous structures are etched on the surface of PEEK after sulfonation treatment.A poly(dopamine)layer is coated on the porous surface of the sulfonated PEEK(SPEEK),which provides anchoring groups for the subsequent deposition of the CS layer.Results show that the CS coating on the porous surface of SPEEK significantly improve the hydrophilicity and biomineralization formation of hydroxyapatite.Compared with PEEK,SPEEK-PDA-CS displays higher bioactivity to promote the proliferation and osteogenic differentiation of rBMSCs,including the increase of ALP activity and formation of calcium nodules,the expression of osteogenic differentiation-related genes.These results are beneficial to extending clinical applications of PEEK-based implants for bone tissue repair and orthopedic surgery.
基金This work was supported by grants from the Nature Science Foundation of China(11572209,11272225).
文摘The present study was designed to investigate the role of estrogen receptorα(ERα)in biaxial tensile strain(BTS)regulated osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells(rBMSCs).rBMSCs were derived fromrats and overexpressed ERα.The rBMSCs were subjected to BTS at 1Hz with a strain of 2%for 4 h per day,3 days,with or without ERαinhibitor ICI 182,780(ICI).Then,bone mineralization was performed by Alizarin Red Staining.The markers of osteogenic differentiation and downstream Wnt3a/β-catenin signaling were detected by western blotting.Results showed that BTS enhanced the osteogenic differentiation of rBMSCs,increased protein expression levels of alkaline phosphatase(ALP),runt-related transcription factor 2(Runx2),collagen type I(Col I)and osteocalcin(OCN),and it increased the protein expression levels of estrogen receptor(ER)α(ERα),Wnt3a,andβ-catenin.BTS The activated Wnt3a/β-catenin signaling pathway induced by BTS was abolished by ICI 182,780(ICI).In addition,overexpressing ERαin rBMSCs promoted the osteogenic differentiation by BTS.Taken together,BTS induced osteogenic differentiation of rBMSCs via the ERαand downstream canonical Wnt3a/β-catenin pathway.
文摘Human mesenchymal stem cells(hMSC)can be expanded in vitro and differentiated towards osteogenic,chondrogenic or adipogenic lineages,making them an attractive source for tissue engineering and regenerative medicine.Chitinase-like-proteins(CLPs)belong to the family 18 glycosyl hydrolases and are believed to play a role in inflammation and tissue remodelling.The aim of this study was to determine the effect of the aminosugar glucosamine on the expression of the CLP YKL-40 during osteogenic differentiation of hMSC.Glucosamine did not affect multipotency of hMSC nor proliferation rate of undifferentiated hMSC.YKL-40 was expressed during both expansion of undifferentiated hMSC and during osteogenic differentiation.A slight but non-significant increase in YKL-40 expression was observed with glucosamine,accompanied by a pH-dependent delay in mineralization.However,glucosamine induced higher expression of osteogenic marker genes.
基金funded by Ollie och Elof Ericssons Stifelse for vetenskaplig forskning,Stiftelsen Lars Hiertas Minne and STINT(Stiftelsen for internationalisering av hogre utbildning och forskning).
文摘Nanotopographical features are found to have significant effects on bone behavior. In the present study, nanoporous aluminas with different pore sizes (20, 100 and 200 nm in diameter), were evaluated for their osteoinductive and drug eluting properties. W20-17 marrow stromal cells were seeded on nanoporous alumina with and without the addition of BMP-2. Although cell proliferation was not affected by pore size, osteogenic differentiation was 200 nm as compared to 20 and 100 nm pores induced higher alkaline phosphatase activity (ALP) and osteocalcin expression levels, thus indicating osteoblastic differentiation. Cell morphology revealed that cells cultured on 20 nm pores adopted a rounded shape, while larger pores (200 nm) elicited an elongated morphology. Furthermore, ALP expression levels were consistently higher on BMP-2 loaded nanoporous alumina surfaces compared to unloaded surfaces, indicating that not only is nanoporous alumina osteoinductive, but also has the potential to be used as a drug eluting bone-implant coating.
文摘Objective To study the effects of combination of bone morphogenetic protein-2 (BMP-2) and strontium chloride on proliferation and osteogenic differentiation of human umbilical cord mesenchymal stem cells(hUCM-SCs)in vitro culture.
基金Supported by National Natural Science Foundation of ChinaNo.81600882 and 81870755+4 种基金China Postdoctoral Science FoundationNo. 2019M663009President Foundation of Nanfang HospitalSouthern Medical UniversityNo.2019B002.
文摘Periodontal diseases are infectious diseases that are characterized by progressive damage to dental support tissue.The major goal of periodontal therapy is to regenerate the periodontium destroyed by periodontal diseases.Human periodontal ligament(PDL)tissue possesses periodontal regenerative properties,and periodontal ligament stem cells(PDLSCs)with the capacity for osteogenic differentiation show strong potential in clinical application for periodontium repair and regeneration.Noncoding RNAs(ncRNAs),which include a substantial portion of poly-A tail mature RNAs,are considered“transcriptional noise.”Recent studies show that ncRNAs play a major role in PDLSC differentiation;therefore,exploring how ncRNAs participate in the osteogenic differentiation of PDLSCs may help to elucidate the underlying mechanism of the osteogenic differentiation of PDLSCs and further shed light on the potential of stem cell transplantation for periodontium regeneration.In this review paper,we discuss the history of PDLSC research and highlight the regulatory mechanism of ncRNAs in the osteogenic differentiation of PDLSCs.
基金Supported by the National Natural Science Foundation of China(81372007)
文摘Objective To observe the expression profiles of osteoblast-related genes in human mesenchymal stem cells(MSCs) derived from bone marrow during osteogenic differentiation. Methods MSCs were induced to differentiate with MSC osteogenic differentiation medium for 7, 14, 21 and 28 days respectively. Alizarin Red staining was used to detect matrix mineralization. Expression of osteoblast-related genes, including osteocalcin, osteopontin, Runt-related transcription factor 2(Runx2), alkaline phosphatase and collagen type Ⅰ, was assessed with quantitative reverse transcription-polymerase chain reaction. Results On day 14 after induction of differentiation, cells were stained positively with Alizarin Red. The expression levels of these genes exhibited an upward trend as induction time was prolonged. Exposure to osteogenic differentiation medium less than 21 days did not significantly induce osteocalcin expression; osteocalcin expression levels in the differentiated cells induced for 21 and 28 days were 1.63 and 2.46 times as high as the undifferentiated cells respectively(all P<0.05). Stimulation with MSC osteogenic differentiation medium over 14 days significantly enhanced bone marrow-derived MSCs to express osteopontin and Runx2 genes(all P<0.05). Osteogenic differentiation medium could significantly induce the expressions of alkaline phosphatase and collagen type Ⅰgenes(all P<0.05). Their expressions reached the peak levels on day 21, which were increased more than 4- and 3-fold respectively. Conclusion Human bone marrow-derived MSCs could exhibit the sequential expression pattern of osteoblast marker genes during osteogenic differentiation in vitro.
基金supported by the Clinic of Oral and Maxillofacial Surgery and the medical faculty of the Georg-August-University Gottingen, Germany
文摘Sites of implantation with compromised biology may be unable to achieve the required level of angiogenic and osteogenic regeneration. The specific function and contribution of different cell types to the formation of prevascularized, osteogenic networks in co-culture remains unclear. To determine how bone marrow-derived mesenchymal stromal cells(BMSCs) and endothelial cells(ECs) contribute to cellular proangiogenic differentiation, we analysed the differentiation of BMSCs and ECs in standardized monolayer, Transwell and co-cultures. BMSCs were derived from the iliac bone marrow of five patients,characterized and differentiated in standardized monolayers, permeable Transwells and co-cultures with human umbilical vein ECs(HUVECs). The expression levels of CD31, von Willebrand factor, osteonectin(ON) and Runx2 were assessed by quantitative reverse transcriptase polymerase chain reaction. The protein expression of alkaline phosphatase, ON and CD31 was demonstrated via histochemical and immunofluorescence analysis. The results showed that BMSCs and HUVECs were able to retain their lineage-specific osteogenic and angiogenic differentiation in direct and indirect co-cultures. In addition, BMSCs demonstrated a supportive expression of angiogenic function in co-culture, while HUVEC was able to improve the expression of osteogenic marker molecules in BMSCs.
基金supported by grants from the Kaohsiung Medical University of Taiwan (KMU-Q099018 and KMU-Q098025)
文摘Retaining or improving periodontal ligament (PDL) function is crucial for restoring periodontal defects. The aim of this study was to evaluate the physiological effects of low-power laser irradiation (LPLI) on the proliferation and osteogenic differentiation of human PDL (hPDL) cells. Cultured hPDL cells were irradiated (660 nm) daily with doses of 0, 1, 2 or 4 J·cm-2 . Cell proliferation was evaluated by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay, and the effect of LPLI on osteogenic differentiation was assessed by Alizarin Red S staining and alkaline phosphatase (ALP) activity. Additionally, osteogenic marker gene expression was confirmed by real-time reverse transcription-polymerase chain reaction (RT-PCR). Our data showed that LPLI at a dose of 2 J·cm-2 significantly promoted hPDL cell proliferation at days 3 and 5. In addition, LPLI at energy doses of 2 and 4 J·cm-2 showed potential osteogenic capacity, as it stimulated ALP activity, calcium deposition, and osteogenic gene expression. We also showed that cyclic adenosine monophosphate (cAMP) is a critical regulator of the LPLI-mediated effects on hPDL cells. This study shows that LPLI can promote the proliferation and osteogenic differentiation of hPDL cells. These results suggest the potential use of LPLI in clinical applications for periodontal tissue regeneration.
基金supported by 2021 Nantong City Basic Research and People's Livelihood Science and Technology Plan Guiding Project (JCZ21133)。
文摘Chronic high glucose(HG) plays a crucial role in the pathogenesis of diabetes-induced osteoporosis by inhibiting the differentiation and proliferation of osteoblasts. This study aims to examine the role of E26 transformation-specific 1(ETS1) in the inhibition of osteoblast differentiation and proliferation caused by chronic HG, as well as the underlying mechanism. Chronic HG treatment downregulated ETS1 expression and inhibited differentiation and proliferation of MC3 T3-E1 cells. Downregulation of ETS1 expression inhibited the differentiation and proliferation of MC3 T3-E1 cells under normal glucose conditions, and ETS1 overexpression attenuated the damage to cells exposed to chronic HG. In addition, ETS1 overexpression reversed the decrease in runt-related transcription factor 2(Runx2) expression in MC3 T3-E1 cells treated with chronic HG. Using chromatin immunoprecipitation(ChIP) and luciferase reporter assays, we confirmed that ETS1 directly bound to and increased the activity of the Runx2 promoter. In summary, our study suggested that ETS1 was involved in the inhibitory effect of chronic HG on osteogenic differentiation and proliferation and may be a potential therapeutic target for diabetes-induced osteoporosis.
基金the National Natural Science Foundation of China (No. 30200063, 30470483)
文摘Poly (lactic acid/glycolic acid/asparagic acid-co-polyethylene glycol)(PLGA-[ASP-PEG]) scaffold materials were linked with a novel nonviral vector (K)16GRGDSPC through cross linker Sulfo- LC-SPDP to construct a new type of nonviral gene transfer system. Eukaryotic expressing vector containing transforming growth factor beta 1 (pcDNA3-TGFβ1) was encapsulated by the system. Bone marrow stromal cells (BMSCs) obtained from rabbit were cultured on PLGA-[ASP-PEG] modified by (K)16GRGDSPC and TGF-β1 gene and PLGA-[ASP-PEG] modified by (K)16GRGDSPC and empty vector pcDNA3 as control. The expressions of osteogenic makers of the BMSCs cultured on the TGF-β1 gene-activated scaffold materials were found significantly higher than those of the control group (P<0.05). A brand-new way was provided for regulating seed cells to directionally differentiate into osteoblasts for bone defect restoration in bone tissue engineering.
基金the Fundamental Research Funds for the Central Universities,China(No.14D110519)Pujiang Talent Program Funded by the Science and Technology Commission of Shanghai Municipality,China(No.10PJ1400200)National Natural Science Foundation of China(No.51073032)
文摘Bone tissue engineering, aiming at developing bone substitutes for repair and regeneration of bone defects instead of using autologous bone grafts,has attracted wide attention in the field of tissue engineering and regenerative medicine.Developing biomimetic biomaterial scaffolds able to regulate osteogenic differentiation of stem cells could be a promising strategy to improve the therapeutic efficacy.In this study, electrospun composite nanofibers of hydroxyapatite / collagen / chitosan( HAp / Col / CTS)resembling the fibrous nanostructure and constituents of the hierarchically organized natural bone,were prepared to investigate their capacity for promoting bone mesenchymal stem cells( BMSCs)to differentiate into the osteogenic lineage in the absence and presence of the osteogenic supplementation, respectively.Cell morphology,proliferation and quantified specific osteogenic protein expression on the electrospun HAp / Col / CTS scaffolds were evaluated in comparison with different controls including electrospun nanofibrous CTS,HAp / CTS and tissue culture plate.Our results showed that the nanofibrous HAp / Col / CTS scaffolds supported better spreading and proliferation of the BMSCs than other substrates( P < 0.01).Expressions of osteogenesis protein markers,alkaline phosphatase( ALP) and Col,were significantly upregulated on the HAp / Col / CTS than those on the CTS( P < 0.01) and HAp /CTS( P < 0.05) scaffolds in the absence of the osteogenic supplementation.Moreover,presence of osteogenic supplementation also proved to enhance osteogenic differentiation of BMSCs on HAp /Col / CTS scaffolds, indicative of a synergistic effect.This study highlights the potential of BMSCs / HAp / Col / CTS cell-scaffold system for functional bone repair and regeneration applications.
文摘The human umbilical cord is a source of numerous Mesenchymal Stem Cells (MSCs), making it as a potential source of allogeneic multipotent cell for bone tissue engineering. The aims of this study were to find: 1) Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) phenotypic characterization, 2) The in-vitro osteogenic differentiation potential of hUCMSCs, 3) The cytotoxicity of gelatin solvent to hUCMSCs using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay. As a result, through characterization of hUCMSCs, the majority of target cells expressed specific MSCs markers, Cellular Differentiation (CD)73, smaller number of subpopulation expressed CD90 with only minimal subpopulation expressed CD105 and all negative MSCs markers. Osteoblastic differentiation was found in a significantly high number of cells when in vitro osteogenic differentiation of hUCMSCs with Alizarin Red staining was done. Biocompatibility analysis using the MTT assay showed that gelatin solvent and Alpha modification of minimum essential medium Eagle (α-MEM) was non-toxic for hUCMSCs in vitro. The study concluded that hUCMSCs isolated from human umbilical cord was capable of undergoing in vitro osteogenesis, indicating its potential as allogeneic stem cells for clinical application in bone tissue engineering.
基金supported by grants from National Natural Science Foundation of China(No.81360279)Guangxi Natural Science Foundation(No.2015GXNSFAA139150)
文摘Objective:To investigate the effects and mechanisms of Polygonatum sibiricum polysaccharide(PSP)on osteogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods:The mouse BMSCs were cultured and induced in osteoblast medium(OBM)containing finalconcentrations(0 mg/L,12.5 mg/L,25 mg/L,and 50 mg/L)of PSP.The proliferation and cytotoxicity of BMSCs were detected by MTT assay.Alkaline phosphatase(ALP)and Alizarin red S staining were performed after 7 days' ossification-inducing culture.The mRNA expressions of ALP,Runx2 and osteocalcin(OCN)were determined by fluorescence quantitative PCR(qPCR).The mRNA and protein expressions of Tafazzin(TAZ)(a key effector of Hippo pathway)were measured by qPCR and western blotting,respectively.Results:PSP was non-cytotoxicwithin the dose range of 12.5-50 mg/L and had no impact on the proliferation of BMSCs.The activity of ALP,the intensity of ALP staining,and the formation of mineralized nodules were increased by PSP treatment(25 and50 mg/L)(P<0.01).Moreover,administration of 25 mg/L PSP significantly enhanced the mRNA levels of osteoblastic differentiation makers ALP,Runx2 and OCN as well as the mRNA and protein expressions of TAZ(P<0.01).Conclusion:PSP could promote osteogenic differentiation of BMSCs,and the mechanisms might be related to the activation of TAZ in the Hippo signaling pathway.
